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* Instituto de Bioquímica (Centro Mixto CSIC-UCM) and
Centro de Citometría de Flujo y Microscopía Confocal, Facultad de Farmacia. Universidad Complutense, 28040 Madrid, Spain
1Correspondence: Instituto de Bioquímica, Facultad de Farmacia, 28040 Madrid. Spain. E-mail: boscal{at}eucmax.sim.ucm.es
Treatment of elicited peritoneal macrophages or the macrophage cell line
RAW 264.7 with high concentrations of nitric oxide donors is followed
by apoptotic cell death. Analysis of the changes in the mitochondrial
transmembrane potential (
m) with specific fluorescent
probes showed a rapid and persistent increase of m, a
potential that usually decreases in cells undergoing apoptosis through
mitochondrial-dependent mechanisms. Using confocal microscopy, the
release of cytochrome c from the mitochondria to the
cytosol was characterized as an early event preceding the rise of
m. The cytochrome c from cells treated
with nitric oxide donors was modified chemically, probably through the
formation of nitrotyrosine residues, suggesting the synthesis of
peroxynitrite in the mitochondria. These results indicate that nitric
oxide-dependent apoptosis in macrophages occurs in the presence of a
sustained increase of m, and that the chemical
modification and release of cytochrome c from the
mitochondria precede the changes of m.—Hortelano,
S., Alvarez, A. M., Boscá, L. Nitric oxide induces tyrosine
nitration and release of cytochrome c preceding an
increase of mitochondrial transmembrane potential in macrophages.
Key Words: apoptosis • NO • nitric oxide synthase • mitochondria
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